Desenvolvimento de forma sólida contendo pré e posbióticos nanoestruturadas para controle da microbiota e proteção cutânea
Data
2024-03-28
Autores
Machado, Ana Carolina Henriques Ribeiro 
Orientadores
Leite-Silva, Vânia Rodrigues
Tipo
Tese de doutorado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Introdução: A antissepsia das mãos, apesar de eficaz de prevenção e propagação de doenças, pode afetar negativamente a função de barreira da pele, desencadeando dermatoses. Frente a esse cenário, recomenda-se o uso de antissépticos para as mãos que possuam também ativos capazes de proteger tanto a barreira física quanto a microbiana, reestabelecendo a sua homeostase. Objetivo: O presente trabalho objetivou desenvolver uma pastilha holobionte, altamente dispersível, com ação reguladora da microbiota, virucida e protetora da barreira da pele. Métodos: Foi realizado um planejamento de experimento fatorial 23 para definir a melhor formulação para o desenvolvimento da pastilha base, mantendo peso médio, dureza, dimensões, taxa de intumescimento e tempo de desintegração como parâmetros a serem analisados. Para a produção das pastilhas holobiontes, a formulação base escolhida foi produzida por compressão direta de matérias-primas prebióticas, posbióticas e excipientes, além de nanopartículas lipídicas sólidas contendo posbióticos, obtidas por homogeneização em alta pressão e liofilizadas. Sua ação virucida in vitro contra partículas de alfa-coronavírus (CCoV - VR809) foi determinada em cultura de células VERO. A análise in vitro, utilizando células em monocamada e pele equivalente humana, foi realizada por reação de transcripitase combinada com reação em cadeia da polimerase (RTq-PCR) em para determinar a expressão de interleucinas 1, 6, 8 e 17, aquaporina-3, involucrina e filagrina, além de forkheadbox O3 e sirtuína-1. As atividades antioxidante e de síntese de colágeno-1 foram realizadas em fibroblastos. Análise metagenômica do microbioma da pele foi determinada in vivo antes e após a aplicação da pastilha holobionte, durante uma semana de uso contínuo e, foram comparadas ao uso do álcool em gel. As amostras foram analisadas avaliando sequenciamento da região V3-V4 do gene 16S rRNA. As características biofísicas da pele do dorso das mãos foram avaliadas in vivo. Teste de Diferença do Controle foi utilizado para estabelecer uma avaliação sensorial discriminativa. Resultados: Após serem definidas como seguras, a avaliação virucida da pastilha obteve resultado igual ou superior ao álcool 70%, com diminuição de interleucinas, e manutenção ou melhora dos marcadores gênicos de barreira da pele, in vitro. Observou-se ainda, nas avaliações in vivo, o reestabelecimento da microbiota e da barreira cutânea após o uso da pastilha holobionte. Já na avaliação sensorial discriminativa, as formulações contendo modificadores de sensorial não se diferenciaram do padrão, sendo consideradas semelhantes. Conclusões: As pastilhas holobiontes foram capazes de melhorar os marcadores gênicos relacionados com a barreira cutânea e a sua microbiota, sendo favorável para o uso como antisséptico em substituição ao uso do álcool 70%.
Introduction: Hand disinfection, despite being effective in preventing and spreading diseases, can negatively affect the skin's barrier function, triggering dermatoses. According to this, it is recommended to use hand antiseptics that also contain active ingredients able to protect both, the physical and microbial barriers, reestablishing their homeostasis. Objective: This study aimed to develop a holobiont tablet, fast dispersibility, with regulation of the microbiota, virucide activity and protective of the skin barrier. Methods: A 23 factorial experiment was planned to define the best formulation for the development of the base tablet, keeping average weight, hardness, dimensions, swelling rate and disintegration time as parameters to be analyzed. To produce holobiont tablets, the chosen base formulation was produced by direct compression of prebiotic, postbiotic and excipient, in addition was used solid lipid nanoparticles containing postbiotics, obtained by high-pressure homogenization and freeze-dried. Virucide activity in vitro, against alpha-coronavirus particles (CCoV - VR809), was determined in VERO cell culture. In vitro analysis, using monolayer cells and human equivalent skin, was performed by reverse transcription polymerase chain reaction RTq-PCR to determine the expression of interleukins 1, 6, 8 and 17, aquaporin-3, involucrin and filaggrin, besides forkheadbox and sirtuin-1. Antioxidant activity and collagen-1 synthesis were also performed in fibroblast cells. Metagenomic analysis of the skin microbiome was determined in vivo before and after application of the holobionte tablet, during one week of continuous use, and compared to the use of alcohol gel. Samples were analyzed by sequencing the V3-V4 region of the 16S rRNA gene. The biophysical characteristics of the skin on the back of the hands were evaluated in vivo. Control Difference Test was used to establish a discriminative sensory evaluation. Results: After being defined as safe, the virucidal evaluation of the tablet obtained results equal to or greater than 70% alcohol, with a reduction in interleukins, and maintenance or improvement of skin barrier gene markers. In addition, in the in vivo assays, there was the reestablishment of the microbiota and skin barrier after use of holobiont tablet. Already, in the sensory analysis, it can also be observed that the formulations containing sensory modifiers did not differ from the standard, being considered similar. Conclusions: The holobiont tablets were able to improve the genetic markers related to the skin barrier and also its microbiota, being favorable for use as a hand sanitizer instead of the use of 70% alcohol 70%.
Introduction: Hand disinfection, despite being effective in preventing and spreading diseases, can negatively affect the skin's barrier function, triggering dermatoses. According to this, it is recommended to use hand antiseptics that also contain active ingredients able to protect both, the physical and microbial barriers, reestablishing their homeostasis. Objective: This study aimed to develop a holobiont tablet, fast dispersibility, with regulation of the microbiota, virucide activity and protective of the skin barrier. Methods: A 23 factorial experiment was planned to define the best formulation for the development of the base tablet, keeping average weight, hardness, dimensions, swelling rate and disintegration time as parameters to be analyzed. To produce holobiont tablets, the chosen base formulation was produced by direct compression of prebiotic, postbiotic and excipient, in addition was used solid lipid nanoparticles containing postbiotics, obtained by high-pressure homogenization and freeze-dried. Virucide activity in vitro, against alpha-coronavirus particles (CCoV - VR809), was determined in VERO cell culture. In vitro analysis, using monolayer cells and human equivalent skin, was performed by reverse transcription polymerase chain reaction RTq-PCR to determine the expression of interleukins 1, 6, 8 and 17, aquaporin-3, involucrin and filaggrin, besides forkheadbox and sirtuin-1. Antioxidant activity and collagen-1 synthesis were also performed in fibroblast cells. Metagenomic analysis of the skin microbiome was determined in vivo before and after application of the holobionte tablet, during one week of continuous use, and compared to the use of alcohol gel. Samples were analyzed by sequencing the V3-V4 region of the 16S rRNA gene. The biophysical characteristics of the skin on the back of the hands were evaluated in vivo. Control Difference Test was used to establish a discriminative sensory evaluation. Results: After being defined as safe, the virucidal evaluation of the tablet obtained results equal to or greater than 70% alcohol, with a reduction in interleukins, and maintenance or improvement of skin barrier gene markers. In addition, in the in vivo assays, there was the reestablishment of the microbiota and skin barrier after use of holobiont tablet. Already, in the sensory analysis, it can also be observed that the formulations containing sensory modifiers did not differ from the standard, being considered similar. Conclusions: The holobiont tablets were able to improve the genetic markers related to the skin barrier and also its microbiota, being favorable for use as a hand sanitizer instead of the use of 70% alcohol 70%.